Journal
EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES
Volume 31, Issue 6, Pages 1073-1079Publisher
SPRINGER
DOI: 10.1007/s10096-011-1409-5
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Funding
- Foundation Idella [3.3.5-2010/II]
- Wellcome Trust
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The purpose of this study was to validate a multiplex real-time PCR assay capable of detecting toxigenic and simultaneously identifying ribotype 027/ST-1 by targeting the toxin genes and in one reaction and in a separate reaction identifying the Delta 117 deletion in associated with ribotype 027/ST-1. PCR was done prospectively on 704 samples routinely submitted to our department and results were compared to results of toxigenic culture. Sequencing of , multi locus sequence typing (MLST) and PCR ribotyping were done on cultured isolates to confirm the correct identification of the Delta 117 deletion in and ribotype 027/ST-1, respectively. The PCR assay displayed a sensitivity, specificity, PPV and NPV of 99.0%, 97.4%, 87.4% and 99.8%, respectively, compared to toxigenic culture on 665 samples evaluable both by PCR and culture. Sequencing of , ribotyping and MLST of cultured isolates validated the genotyping assay and confirmed the ability of the assay to correctly identify ribotype 027/ST-1 in our current epidemiological setting. We describe the use of a combination of two separate PCR assays for sensitive and specific detection of toxigenic and presumptive identification of C. difficile 027/ST-1.
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